Pinostrobin targets the PI3K/AKT/CCL2 axis in intestinal epithelial cells to inhibit intestinal macrophage infiltration and alleviate dextran sulfate sodium-induced colitis in mice.
10.12122/j.issn.1673-4254.2025.10.16
- Author:
Keni ZHANG
1
;
Tong QIAO
1
;
Lin YIN
1
;
Ju HUANG
2
;
Zhijun GENG
2
;
Lugen ZUO
3
;
Jianguo HU
1
;
Jing LI
1
Author Information
1. Department of Clinical Laboratory,First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China.
2. Central Laboratory,First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China.
3. Anhui Provincial Key Laboratory of Basic and Translational Research on Inflammatory Diseases, First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China.
- Publication Type:Journal Article
- Keywords:
CCL 2;
PI3K/AKT signaling pathway;
colitis;
dextran sulfate sodium;
infiltration of macrophages;
pinostrobin
- MeSH:
Animals;
Mice;
Mice, Inbred C57BL;
Phosphatidylinositol 3-Kinases/metabolism*;
Colitis/drug therapy*;
Dextran Sulfate;
Proto-Oncogene Proteins c-akt/metabolism*;
Macrophages;
Chemokine CCL2/metabolism*;
Humans;
Signal Transduction/drug effects*;
Caco-2 Cells;
RAW 264.7 Cells;
Epithelial Cells/drug effects*;
Intestinal Mucosa/metabolism*
- From:
Journal of Southern Medical University
2025;45(10):2199-2209
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:To investigate the mechanism through which pinostrobin (PSB) alleviates dextran sulfate sodium (DSS)-induced colitis in mice.
METHODS:C57BL/6 mice were randomized into control group, DSS model group, and PSB intervention (30, 60, and 120 mg/kg) groups. Colitis severity of the mice was assessed by examining body weight changes, disease activity index (DAI), colon length, and histopathology. The expressions of tight junction proteins ZO-1 and claudin-1 in the colon tissues were examined using immunofluorescence staining, and macrophage infiltration and polarization were analyzed with flow cytometry. ELISA and RT-qPCR were used for detecting the expressions of inflammatory factors (TNF‑α and IL-6) and chemokines (CCL2, CXCL10, and CX3CL1) in the colon tissues, and PI3K/AKT phosphorylation levels were analyzed with Western blotting. In cultured Caco-2 and RAW264.7 cells, the effect of PSB on CCL2-mediated macrophage migration was assessed using Transwell assay. Network pharmacology analysis was performed to predict the key pathways that mediate the therapeutic effect of PSB.
RESULTS:In DSS-induced mouse models, PSB at 60 mg/kg optimally alleviated colitis, shown by reduced weight loss and DAI scores and increased colon length. PSB treatment significantly upregulated ZO-1 and claudin-1 expressions in the colon tissues, inhibited colonic macrophage infiltration, and promoted the shift of macrophage polarization from M1 to M2 type. In cultured intestinal epithelial cells, PSB significantly inhibited PI3K/AKT phosphorylation and suppressed chemokine CCL2 expression. PSB treatment obviously blocked CCL2-mediated macrophage migration of RAW264.7 cells, which could be reversed by exogenous CCL2. Network pharmacology analysis and rescue experiments confirmed PI3K/AKT and CCL2 signaling as the core targets of PSB.
CONCLUSIONS:PSB alleviates DSS-induced colitis in mice by targeting intestinal epithelial PI3K/AKT signaling, reducing CCL2 secretion, and blocking macrophage chemotaxis and migration, highlighting the potential of PSB as a novel natural compound for treatment of inflammatory bowel disease.
